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Development of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) thermal inactivation method with preservation of diagnostic sensitivity

Journal of Microbiology 2020년 58권 10호 p.886 ~ 891
김영일, Casel Mark Anthony B., 김세미, 김성규, 박수진, 김은하, 정혜원, 부하령, 최영길,
소속 상세정보
김영일 ( Kim Young-Il ) - Chungbuk National University College of Medicine Medical Research Institute
 ( Casel Mark Anthony B. ) - Chungbuk National University College of Medicine Medical Research Institute
김세미 ( Kim Se-Mi ) - Chungbuk National University College of Medicine Medical Research Institute
김성규 ( Kim Seong-Gyu ) - Chungbuk National University College of Medicine Medical Research Institute
박수진 ( Park Su-Jin ) - Chungbuk National University College of Medicine Medical Research Institute
김은하 ( Kim Eun-Ha ) - Chungbuk National University College of Medicine Medical Research Institute
정혜원 ( Jeong Hye-Won ) - Chungbuk National University College of Medicine Medical Research Institute
부하령 ( Poo Ha-Ryoung ) - Korea Research Institute of Bioscience and Biotechnology Infectious Disease Research Center
최영길 ( Choi Young-Ki ) - Chungbuk National University College of Medicine Medical Research Institute

Abstract


Various treatments and agents had been reported to inactivate RNA viruses. Of these, thermal inactivation is generally considered an effective and cheap method of sample preparation for downstream assays. The purpose of this study is to establish a safe inactivation method for SARS-CoV-2 without compromising the amount of amplifiable viral genome necessary for clinical diagnoses. In this study, we demonstrate the infectivity and genomic stability of SARSCoV- 2 by thermal inactivation at both 56°C and 65°C. The results substantiate that viable SARS-CoV-2 is readily inactivated when incubated at 56°C for 30 min or at 65°C for 10 min. qRT-PCR of specimens heat-inactivated at 56°C for 30 min or 65°C for 15 min revealed similar genomic RNA stability compared with non-heat inactivated specimens. Further, we demonstrate that 30 min of thermal inactivation at 56°C could inactivate viable viruses from clinical COVID-19 specimens without attenuating the qRT-PCR diagnostic sensitivity. Heat treatment of clinical specimens from COVID-19 patients at 56°C for 30 min or 65°C for 15 min could be a useful method for the inactivation of a highly contagious agent, SARS-CoV-2. Use of this method would reduce the potential for secondary infections in BSL2 conditions during diagnostic procedures. Importantly, infectious virus can be inactivated in clinical specimens without compromising the sensitivity of the diagnostic RT-PCR assay.

키워드

SARS-CoV-2; heat inactivation; COVID-19; genomic stability; RT-PCR

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